1. Multi-site assessment of the precision and reproducibility of multiple reaction monitoring–based measurements of proteins in plasma
- Author
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Helene L. Cardasis, Lei Zhao, Angela M. Jackson, Christopher R. Kinsinger, Richard K. Niles, Jason M. Held, Steven J. Skates, Asokan Mulayath Variyath, Susan E. Abbatiello, David L. Tabb, Daniel C. Liebler, Charles Buck, Tara Hiltke, Mu Wang, Paul A. Rudnick, Terri A. Addona, Jeffrey R. Whiteaker, Ronald K. Blackman, Amanda G. Paulovich, Derek Smith, D. R. Mani, Trenton C. Pulsipher, Lorenzo Vega-Montoto, Mehdi Mesri, Bradford W. Gibson, Asa Wahlander, Michael P. Cusack, David F. Ransohoff, Christoph H. Borchers, Eric B. Johansen, Susan J. Fisher, Simon Allen, Clifford H. Spiegelman, Henry Rodriguez, Steven A. Carr, David M. Bunk, Paul Tempst, Nathan G. Dodder, Fred E. Regnier, Sofia Waldemarson, Hasmik Keshishian, Thomas A. Neubert, N. Leigh Anderson, Steven C. Hall, Birgit Schilling, Jing Li, Tony J. Tegeler, Amy-Joan L. Ham, and Lisa J. Zimmerman
- Subjects
Technology Assessment, Biomedical ,Proteome ,Protein biomarkers ,Biomedical Engineering ,Bioengineering ,Computational biology ,Biology ,Bioinformatics ,Proteomics ,Sensitivity and Specificity ,Applied Microbiology and Biotechnology ,Mass Spectrometry ,Humans ,Biomarker discovery ,Detection limit ,Reproducibility ,Selected reaction monitoring ,Multi site ,Reproducibility of Results ,Blood Proteins ,Targeted mass spectrometry ,Linear Models ,Molecular Medicine ,Biomarkers ,Blood Chemical Analysis ,Biotechnology - Abstract
Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the discovery pipeline that bridges unbiased biomarker discovery to preclinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry can quantify candidate protein biomarkers in plasma, reproducibility and transferability of these assays between laboratories have not been demonstrated. We describe a multilaboratory study to assess reproducibility, recovery, linear dynamic range and limits of detection and quantification of multiplexed, MRM-based assays, conducted by NCI-CPTAC. Using common materials and standardized protocols, we demonstrate that these assays can be highly reproducible within and across laboratories and instrument platforms, and are sensitive to low mug/ml protein concentrations in unfractionated plasma. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma.
- Published
- 2009